Methods for the treatment of glioblastoma multiforme

ABSTRACT

Methods for the treatment of cancer, including glioblastoma multiforme (GBM) and recurrent glioblastoma multiforme (RGBM). The method includes administering to a patient a plurality of extraneous antineoplastons and a plurality of targeted agents. The plurality of extraneous antineoplastons may include two or more antineoplastons selected from phenylacetylglutaminate sodium (PG), phenylacetylisoglutaminate sodium (iso-PG), and phenylacetate (PN). The plurality of targeted agents may include two or more agents selected from bevacizumab, pazopanib, sorafenib, dasatinib, and everolimus.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation application of U.S. Ser. No.15/589,887, filed May 8, 2017, the disclosure of which is incorporatedherein by reference its entirety.

FIELD OF TECHNOLOGY

The present disclosure is directed to methods of treating cancer. Thepresent disclosure is further directed to methods for the treatment ofglioblastoma and recurrent glioblastoma using antineoplastons andtargeted agents.

BACKGROUND

Glioblastoma multiforme (GBM) represents the most frequently occurringand among the most aggressive form of primary malignant brain tumors,accounting for 54% of glioma. A sub-analysis in an internationalrandomized trial by the European Organization for Research and Treatmentof Cancer (EORTC) and the National Cancer Institute of Canada (NCIC)compared the results of radiation therapy (RT) alone with those ofconcomitant RT and temozolomide (TMZ) and found that the addition of TMZto RT for newly diagnosed GBM resulted in a significant survivalbenefit. A significant survival benefit of TMZ administration was alsofound in the subgroup analysis of the 5-year survival data of theEORTC/NCIC trial. Since then, TMZ has been the first-linechemotherapeutic agent for GBM. However, despite aggressive treatmentincluding surgery, radiation therapy (RT), and adjuvant TMZ-basedchemotherapy, the prognosis of patients with GBM has been extremelypoor, with a median survival of 14.6 months from diagnosis. The medianprogression-free-survival (PFS) after standard therapy is 6-9 months,demonstrating that current treatment options are palliative rather thancurative.

The Cancer Genome Atlas Network has cataloged recurrent genomicabnormalities in GBM. The Network described a gene expression-basedmolecular classification of GBM into Proneural, Neural, Classical, andMesenchymal subtypes and integrated multidimensional genomic data toestablish patterns of somatic mutations and DNA copy number. Aberrationsand gene expression of EGFR, NF1, PDGFRA/IDH1, and EGFR/neuron markerswere used to define the Classical, Mesenchymal, Proneural, and Neuralsubtypes, respectively. The response of GBM to therapy has been found todiffer by subtype.

The prognosis for recurrent glioblastoma (RGBM) is much worse than forGBM, with a majority of patients dying within six months. In general,treatment of RGBM may involve repeated resection, focal RT, chemotherapyor experimental therapies. However, there has been very modest progressin the treatment of RGBM and there are no standard treatmentrecommendations for RGBM. In view of the unfavorable survival outlookwith currently available treatment modalities, new methods for thetreatment for GBM and RGBM are desirable.

BRIEF DESCRIPTION OF THE DRAWINGS

Embodiments of the present application are described, by way of exampleonly, with reference to the attached Figures, wherein:

FIG. 1 is an illustration depicting a proposed mechanism of action ofantineoplaston (ANP)/phenylbutyrate (PB) therapy in RGBM, according toan example embodiment of the present disclosure;

FIG. 2 is an illustration depicting the inhibition of growth, survival,invasion, angiogenesis, and cell cycle maintenance by ANP/PB therapy incombination with targeted agents, according to an example embodiment ofthe present disclosure; and

FIG. 3 is an illustration depicting the inhibition of signalingpathways, metabolism, drug resistance, and cell cycle maintenance andpromotion of apoptosis by ANP/PB therapy in combination with targetedagents, according to an example embodiment of the present disclosure.

It should be understood that the various aspects are not limited to thedepictions provided in the drawings.

DETAILED DESCRIPTION

Various embodiments of the disclosure are discussed in detail below.While specific implementations are discussed, it should be understoodthat this is done for illustration purposes only. A person skilled inthe relevant art will recognize that other procedures and techniques canbe used without parting from the spirit and scope of the presentdisclosure.

It should be understood at the outset that although illustrativeimplementations of one or more embodiments are illustrated below, thedisclosed methods can be implemented using any number of techniques. Thedisclosure should in no way be limited to the illustrativeimplementations, drawings, and techniques disclosed herein, but can bemodified within the scope of the appended claims along with their fullscope of equivalents. In addition, numerous specific details are setforth in order to provide a thorough understanding of the embodimentsdescribed herein. However, it will be understood by those of ordinaryskill in the art that the embodiments described herein can be practicedwithout these specific details. In other instances, methods, proceduresand techniques have not been described in detail so as not to obscurethe related relevant feature being described.

The following definitions are provided in order to aid those skilled inthe art in understanding the present disclosure. As used herein, theterm “objective response” refers to a response of a tumor to atherapeutic treatment. In at least some instances, the term “objectiveresponse” refers to a measured reduction in tumor size. In otherinstances, the term “objective response” can also refer a stopping orarresting of the growth of the tumor. As used herein, the term“concurrently” as in “concurrent administration,” in all its forms anduses, refers to the overlapping administration of one or moretherapeutic compositions or agents on the same day during at least aportion of its therapeutic regimen. Therefore, for example, the dailyintravenous administration of a therapeutic agent administered on atleast one overlapping day that a second therapeutic composition isadministered orally would represent concurrent administration.

Further, a therapeutic composition administered intravenously once everytwo weeks is concurrently administered with a therapeutic compositionthat is orally administered daily if the intravenous administrationoccurs on the same day as at least one day that the oral administrationoccurs. As used herein, the term “antineoplaston (ANP) therapy,” refersto administration to a patient, by any administration route, of an “ANPtherapeutic composition” comprising a therapeutically effective amountof Atengenal (A10), Astugenal (AS2-1), or any combination thereof.

The present disclosure provides methods for treating cancer in apatient. According to at least one aspect of the present disclosure, themethod includes administering to a patient a plurality of extraneousantineoplastons and a plurality of targeted agents. The plurality oftargeted agents may include pazopanib, sorafenib, dasatinib, everolimus,and bevacizumab. The plurality of extraneous antineoplastons may includephenylacetate (PN), phenylacetylglutaminate sodium (PG),phenylacetylisoglutaminate sodium (iso-PG), and any combination thereof.In at least some instances, the cancer may be glioma. In otherinstances, the cancer may be glioblastoma multiforme (GBM) or recurrentglioblastoma multiforme (RGBM).

In at least some instances, the presently disclosed methods may be usedto treat a patient that has failed treatment with surgery, radiationtherapy and/or chemotherapy prior to administration of the plurality ofextraneous antineoplastons and the plurality of targeted agents. Forinstance, the presently disclosed method may include treating a patientthat has previously failed to achieve an objective response followingadministration of chemotherapy with temozolomide (TMZ) and/orbevacizumab (BVZ). In some instances, the presently disclosed methodsmay be used to treat a patient that has previously undergone a resectionof a glioma prior to administration of the plurality of extraneousantineoplastons and the plurality of targeted agents. According to atleast one aspect of the present disclosure, the method may furtherinclude genomic profiling of a tumor in the patient to determine thegenetic subtype. In at least some instances, the method may includetreating GBM or RGBM in a patient in which the GBM or RGBM is of theClassical or Proneural subtype.

Antineoplastons (ANP) are peptides, amino acid derivatives andcarboxylic acids which were initially isolated from the blood and urineof healthy subjects. Because ANP were found to be deficient in the bloodof cancer patients, it was postulated that they may have anticanceractivity. Antineoplastic activity of these compounds has been shown in anumber of clinical and preclinical studies. ANP may be administered to apatient in the form of an ANP therapeutic composition that includesAtengenal (A10), Astugenal (AS2-1), or any combination thereof.

Atengenal (A10) comprises a 4:1 ratio of syntheticphenylacetylglutaminate sodium (PG) and phenylacetylisoglutaminatesodium (iso-PG). PG has a molecular weight of 286.26 and an empiricalformula of C₁₃H₁₅N₂NaO₄. PG was first described by Thierfelder andSherwin and is synthesized by the reaction of phenylacetyl chloride withL-glutamine in an aqueous solution containing sodium bicarbonate. PG isa hygroscopic white powder having a melting point of approximately 102°C. and is very soluble in water. The structural formula of PG is:

Iso-PG has a molecular weight of 286.26 and an empirical formula ofC₁₃H₁₅N₂NaO₄. Iso-PG is synthesized by the reaction of phenylacetylchloride with L-glutamine in an aqueous solution containing sodiumbicarbonate to afford PG, which in turn is heated under vacuum at 160°C. to yield A10C (3-phenylacetylamino-2,6-piperidinedione). When A10C istreated with sodium hydroxide, it produces a mixture of PG and iso-PG ina 4:1 ratio. Iso-PG is a white powder having a melting point ofapproximately 175-176° C. and is soluble in water. The structuralformula of iso-PG is:

Astugenal (AS2-1) comprises phenylacetate (PN) and PG in a 4:1 ratio. PNis characterized by a molecular weight of 158.63 and an empiricalformula of C₈H₈NaO₂. PN is synthesized by refluxing benzyl cyanide withdilute sulfuric acid or hydrochloric acid. In solid form, PN has amelting point of approximately 76.5° C. The structural formula of PN is:

In at least some instances, ANP therapeutic compositions may beadministered to a patient intravenously using, for instance, adual-channel infusion pump and central venous catheter. In some cases,intravenous administration of ANP therapeutic compositions may occuronce every four hours at the infusion rate of from about 50 mL/hr toabout 250 mL/hr depending on patient's age and condition/tolerance, inan outpatient setting. AS2-1 or A10 can be administered separately incombination with targeted agents. In such cases, a single channel pumpcan be used. They can also be administered every 5, 6, or 8 hours.

A published study evaluated the safety and efficacy of ANP therapy inrecurrent high-grade glioma, with a special emphasis on RGBM. Burzynskiet al. (2014) A phase II study of antineoplastons A10 and AS2-1 in adultpatients with recurrent glioblastoma multiforme: Final report (protocolBT-21). J. Cancer Ther 5:946-956. In twenty-four patients with RGBM whoreceived at least four weeks of ANP therapy, four patients (16.7%)achieved an objective response while overall survival was 39.3% at oneyear and 4.4% at two, five, and ten years. A group of RGBM patients withunusually long survival following ANP therapy were identified. Burzynskiet al. (2014) Recurrent glioblastoma multiforme—A strategy for long-termsurvival. J. Cancer Ther 5:957-976. Tumor tissue from one of thesepatients, who was alive and free of disease 11 years after ANP therapyfor RGBM, was collected for gene expression profiling. Burzynski et al.(2015) Comprehensive genomic profiling of recurrent classic glioblastomain a patient surviving eleven years following antineoplaston therapy.Cancer Clin. Oncol. 4(2):41-52. In December 2014, comprehensive genomicprofiling of the patient tumor tissue was performed by FoundationMedicine, Inc. of Cambridge, Mass., using a Next Generation Sequencing(NGS) assay of 343 cancer-related genes and introns. Based on the NGSassay, the patient's RGBM was classified as the Classical subtype. Theresponse of GBM to ANP therapy has been found to differ by subtype, withthe greatest benefit being seen in the Classical and Proneural subtypes,which generally responds the least of all the subtypes to standardtherapy.

Published data indicate that only some GBM patients will respond to ANPtherapy when administered alone. Burzynski et al. (2014) Recurrentglioblastoma multiforme—A strategy for long-term survival. J. CancerTher 5:957-976. While PG and PN affect over 100 abnormal genes in theGBM genome, their effect, taken alone, may not be sufficient toconsistently alter the GBM genome (˜650 abnormal genes) and produce anobjective response. Therefore, the present disclosure provides methodsfor the treatment of cancer, including GBM and RGBM, that includes acombination of therapeutic agents.

RGBM patients have been treated with a combination of sodiumphenylbutyrate (PB) and additional targeted agents in conjunction withgenomic profiling. Burzynski et al. (2014) Preliminary findings on theUse of Targeted Therapy with Pazopanib and Other Agents in Combinationwith Sodium Phenylbutyrate in the Treatment of Glioblastoma Multiform.J. Cancer Ther 5:1423-1437. Based on this treatment approach, objectiveresponse rates and overall survival exceeded the previously publishedresults in RGBM. Burzynski et al. (2014) J. Cancer Ther 5:946-956.Eleven RGBM patients treated with PB in combination with bevacizumab,dasatinib, pazopanib, and m-TOR inhibitors resulted in six patients(54.5%) achieving an objective response and three patients maintainingstable disease (27.3%), providing for an overall disease control rate of81.8%. The most effective therapeutic regime consisted of PB,bevacizumab, dasatinib, everolimus and pazopanib. All patients whocomplied with this treatment regime achieved an objective response.Burzynski et al. (2014) J. Cancer Ther 5:1423-1437.

PB is a histone deacetylase inhibitor which is FDA-approved for ureacycle disorders. PB has also been used for the treatment of glioma andacute promyelocytic leukemia. The structural formula of PB is:

The metabolites of PB include PG and PN, the active ingredients in theANP therapy and the ANP therapeutic composition. Without being held toany particular theory regarding the mechanism by which ANP and PBprovides an effective therapeutic method of treating cancer, a proposedmechanism of action of ANP/PB therapy in RGBM is shown in FIG. 1. Theinhibition of growth, survival, invasion, angiogenesis, and cell cyclemaintenance by ANP/PB therapy in combination with targeted agents isshown in FIG. 2. The inhibition of signaling pathways, metabolism, drugresistance, and cell cycle maintenance and promotion of apoptosis byANP/PB therapy in combination with targeted agents is shown in FIG. 3.

While the metabolites of PB include PG and PN, administration of PB maynot be as effective as ANP therapy since PB relies on the generation ofintermediates and products of metabolism, does not contain or produceiso-PG, and because PB is administered orally rather than intravenously.

The present disclosure provides combination therapeutic methods for thetreatment of cancer in a patient. The method includes administering to apatient a plurality of extraneous antineoplastons and a plurality oftargeted agents. In at least some instances, the plurality orantineoplastons and the plurality of targeted agents are administered tothe patient concurrently. Concurrent administration of a plurality ofextraneous antineoplastons and a plurality of targeted agents, accordingto the presently disclosed methods, have been discovered to betherapeutically effective in the treatment of cancer, in particular, inthe treatment of gliomas including GMB and RGMB. Additionally,administration of a plurality of extraneous antineoplastons and aplurality of targeted agents, according to the presently discloseddosage ranges and regimens, has been discovered to be therapeuticallyeffective in the treatment of GMB and RGMB.

The plurality of targeted agents may include two or more agents selectedfrom the group consisting of bevacizumab, pazopanib, sorafenib,dasatinib, and everolimus. Pazopanib and/or sorafenib may be orallyadministered to a patient at a dose of from about 1 mg/kg/day to about12 mg/kg/day. In other cases, pazopanib and/or sorafenib may be orallyadministered to a patient at a dose of from about 2 mg/kg/day to about 6mg/kg/day. In at least some instances, an optimal dose of pazopaniband/or sorafenib may be 3 mg/kg/day.

Dasatinib may be orally administered to a patient at a dose of fromabout 0.3 mg/kg/day to about 2.0 mg/kg/day. In other cases, dasatinibmay be orally administered to a patient at a dose of from about 0.7mg/kg/day to about 1.4 mg/kg/day. In at least some instances, an optimaldose of dasatinib may be 0.7 mg/kg/day.

Everolimus may be orally administered to a patient at a dose of fromabout 0.03 mg/kg/day to about 0.15 mg/kg/day. In other cases, everolimusmay be orally administered to a patient at a dose of from about 0.03mg/kg/day to about 0.10 mg/kg/day. In at least some instances, anoptimal dose of everolimus may be 0.07 mg/kg/day.

Bevacizumab may be administered intravenously to a patient every 1 to 3weeks at a dose of from about 2 mg/kg/day to about 15 mg/kg/day. Inother cases, bevacizumab may be intravenously administered to a patientevery 1 to 3 weeks at a dose of from about 5 mg/kg/day to about 12mg/kg/day. In at least some instances, bevacizumab is optimallyadministered intravenously to a patient every 2 weeks and with anoptimal dose of 10 mg/kg/day.

The plurality of extraneous antineoplastons may be administeredintravenously to a patient. In some instances, the plurality ofextraneous antineoplastons may include phenylacetylglutaminate sodium(PG) and phenylacetylisoglutaminate sodium (iso-PG). For example, theplurality of extraneous antineoplastons may be the Atengenal (A10)composition describe above. In such instances, the plurality ofextraneous antineoplastons may include about a 4:1 ratio of PG andiso-PG. In at least some instances, the plurality of extraneousantineoplastons comprising PG and iso-PG may be administeredintravenously to a patient at a dosage of from about 0.5 g/kg/day toabout 25 g/kg/day. In other instances, the plurality of extraneousantineoplastons comprising PG and iso-PG may be administeredintravenously to a patient at a dosage of from about 2 g/kg/day to about8 g/kg/day. In at least some instances the optimal dose of extraneousantineoplastons comprising PG and iso-PG may be about 4 g/kg/day.

The plurality of extraneous antineoplastons may be administeredintravenously to a patient at a dosage of from about 0.4 g/kg/day toabout 20 g/kg/day phenylacetylglutaminate sodium (PG) and from about 0.1g/kg/day to about 5 g/kg/day phenylacetylisoglutaminate sodium (iso-PG).In other instances, the plurality of extraneous antineoplastons may beadministered intravenously to a patient at a dosage of from about 1.6g/kg/day to about 6.4 g/kg/day phenylacetylglutaminate sodium (PG) andfrom about 0.4 g/kg/day to about 1.6 g/kg/day phenylacetylisoglutaminatesodium (iso-PG). In at least some instances, the optimal dose ofphenylacetylglutaminate sodium (PG) may be 3.2 g/kg/day and the optimaldose of phenylacetylisoglutaminate sodium (iso-PG) may be 0.8 g/kg/day.

In some instances, the plurality of extraneous antineoplastons mayinclude phenylacetate (PN) and phenylacetylglutaminate (PG). Forexample, the plurality of extraneous antineoplastons may be theAstugenal (AS2-1) composition described above. In such instances, theplurality of extraneous antineoplastons may include about a 4:1 ratio ofPN and PG. In at least some instances, the plurality of extraneousantineoplastons comprising PN and PG may be administered intravenouslyto a patient at a dosage of from about 0.04 g/kg/day to about 0.6g/kg/day. In other instances, the plurality of extraneousantineoplastons comprising PN and PG may be administered intravenouslyto a patient at a dosage of from about 0.2 g/kg/day to about 0.4g/kg/day. In at least some instances the optimal dose of extraneousantineoplastons comprising PN and PG may be about 0.4 g/kg/day.

The plurality of extraneous antineoplastons may be administeredintravenously to a patient at a dosage of from about 0.064 g/kg/day toabout 0.48 g/kg/day phenylacetate (PN) and from about 0.016 g/kg/day toabout 0.12 g/kg/day phenylacetylglutaminate sodium (PG). In otherinstances, the plurality of extraneous antineoplastons may beadministered intravenously to a patient at a dosage of from about 0.16g/kg/day to about 0.48 g/kg/day phenylacetate (PN) and from about 0.04g/kg/day to about 0.12 g/kg/day phenylacetylglutaminate sodium (PG). Inat least some instances, the optimal dose of phenylacetate (PN) may be0.32 g/kg/day and the optimal dose of phenylacetylglutaminate sodium(PG) may be 0.08 g/kg/day.

In at least some instances, the plurality of extraneous antineoplastonsmay include phenylacetylglutaminate sodium (PG),phenylacetylisoglutaminate sodium (iso-PG), phenylacetate (PN), and anycombination thereof.

The plurality of extraneous antineoplastons and the pazopanib,sorafenib, dasatinib, and everolimus may be administered daily for oneto seven days. In some instances, the plurality of extraneousantineoplastons and the pazopanib, sorafenib, dasatinib, and everolimusmay be administered daily for at least 8 weeks and the bevacizumab maybe administered intravenously every 1 to 3 weeks for at least 8 weeks.

In at least some instances, the concurrent administration of theplurality of extraneous antineoplastons and the plurality of targetedagents may begin on different days at the beginning of the treatmentregimen in order to better monitor the onset of adverse events. Forexample, the ANP therapy may begin on day 1 and continue daily for theduration of the treatment regimen. On day 2, one of the plurality oftargeted agents may be orally administered and continued daily for theduration of the treatment regimen. On day 3, a second one of theplurality of targeted agents may be orally administered and continueddaily for the duration of the treatment regimen. A third and fourthtargeted agents may be orally administered on day 3 and day 4,respectively, and continued daily for the duration of the treatmentregimen. Intravenous administration of bevacizumab may begin on day 5 ofthe treatment regimen and repeated once every 1-3 weeks thereafter forso long as the treatment regimen continues. No rest period is requiredfor the any of the administered therapeutic compositions. The treatmentregimen may continue, for example, for 8 weeks. An MRI scan may then beused to determine if an objective response has occurred. The treatmentregimen may then be repeated as needed, with or without the staggeredonset of concurrent administration.

In other instances, the ANP therapy may begin on day 1 with escalatingdosages administered on days 2-4. ANP therapy may then continue dailyfor the duration of the treatment regimen. Intravenous administration ofbevacizumab may begin on day 8 of the treatment regimen and repeatedonce every 1-3 weeks thereafter for so long as the treatment regimencontinues. Dasatinib may be administered on day 12 and continued dailyfor the duration of the treatment regimen. Everolimus may beadministered on day 16 and continued daily for the duration of thetreatment regimen. Pazopanib may be administered on day 20 and continueddaily for the duration of the treatment regimen. The treatment regimenmay continue, for example, for 8 weeks. An MRI scan may then be used todetermine if an objective response has occurred. The treatment regimenmay then be repeated as needed, with or without the staggered onset ofconcurrent administration.

According to at least one aspect of the present disclosure, theantineoplaston (ANP) therapy may be administered to a patient incombination with sodium phenylbutyrate (PB). In some instances, the ANPtherapy and/or the intravenous administration of the plurality ofantineoplastons may be replaced by administration of PB.

EXAMPLES Example 1

A 47 year-old male patient from Australia had undergone subtotalresection of a right parieto-occipital GBM followed by treatment withtemozolomide (TMZ) and concomitant radiation therapy. A post-treatmentMRI indicated RGBM. The patient began ANP therapy in combination with aplurality of targeted agents. The patient was intravenously administeredan ANP therapeutic composition comprising 6.3 g/kg/day of A10 and 0.2g/kg/day AS2-1 for approximately 8 months. Concurrently, the patientreceived oral administration of dasatinib (0.5 mg/kg/day), everolimus(0.1 mg/kg/day), and pazopanib (2.1 mg/kg/day) at doses representing a50-80% dose reduction as compared to the standard dose. The patientconcurrently received intravenous administration of bevacizumab (7.5mg/kg) every two weeks. After approximately 26 days of treatment, an MRIrevealed no measureable enhancing disease. A subsequent MRI after 54days of treatment indicated a disappearance of all enhancing disease.After approximately 84 days of treatment, a PET/CT scan revealed nometabolically active disease. The patient returned to Australia after 55days. The patient's current condition is not known.

Example 2

A male patient from Australia had undergone resection of a GBM, whichinvolved his right temporal, parietal, and occipital lobes and thecorpus callosum, followed by treatment with temozolomide (TMZ) andconcomitant radiation therapy. Two weeks after completion of thattherapy, the patient underwent a second surgical resection for RGBM.Subsequently, a MRI revealed a 5.2 cm×4.3 cm tumor. Assisted bymolecular profiling, the patient was administered PB, bevacizumab,erlotinib, rapamycin, and TMZ. After six weeks of treatment there was noevidence of tumor on a repeat MRI scan. The patient remained free ofdisease for over three years when a MRI revealed RGBM. The patientunderwent resection of the RGBM, however, approximately one month later,a MRI revealed a 4.8 cm² right temporal enhancing mass.

At the age of 57, the patient began ANP therapy in combination with aplurality of targeted agents. The patient was intravenously administeredan ANP therapeutic composition comprising 6.6 g/kg/day of A10 and 0.2g/kg/day of AS2-1 for 67 days. Concurrently, the patient received oraladministration of dasatinib (0.5 mg/kg/day), everolimus (0.05mg/kg/day), and pazopanib (2.2 mg/kg/day) at doses representing a 50-80%dose reduction as compared to the standard dose. The patientconcurrently received intravenous administration of bevacizumab (10mg/kg) every two weeks. After approximately 74 days of treatment, a MRIperformed on the patient revealed no measureable enhancing disease. Asubsequent PET/CT scan after 105 days from the start of treatmentrevealed no metabolically active disease. The patient returned toAustralia where is oncologist discontinued treatment. He diedapproximately one year from treatment start.

Example 3

A 23 year-old female patient had been diagnosed with fibrillaryastrocytoma, grade II, IDH1(−). The patient underwent resection of thetumor but developed tumor recurrence within approximately one year, asindicated by an MRI. Further progression was shown by an MRI performedapproximately one month later. The patient underwent a second tumorresection approximately two months later and the pathology showed aglioblastoma, unmethylated MGMT, wild type IDH1 and BRAF mutation. Thepatient received adjuvant radiation therapy for approximately 42 days.Fourteen days later, the patient received oral administration of PB (0.2g/kg/day), sorafenib (3.6 mg/kg/day), everolimus (0.09 mg/kg/day),dasatinib (0.9 mg/kg/day), concurrent with intravenous administration ofbevacizumab (10 mg/kg) every two weeks. A pretreatment MRI revealed acontrast-enhancing lesion measuring 2.5 cm×2.0 cm. A follow-up MRIapproximately 3 months after treatment began indicated no recurrenttumor and a subsequent MRI approximately 46 days later confirmed acomplete response. Two days later, ANP therapy comprising intravenousadministration of AS2-1 through a portable infusion pump wereconcurrently added to the treatment of the patient. The dose of theAS2-1 ANP therapy was gradually increased to 40 mL (80 mg/mL) six timesdaily at 250 mL/hour (0.34 g/kg/day). A follow-up MRI approximately 3months later indicated continuation of the complete response. Thepatient continued the treatment for 8 months. Currently, she is offtreatment for 5 months and continues to be tumor-free.

Example 4

A 71 year-old female patient with a history of hypothyroidism anddiabetes was in her usual state of health until she began having visualimpairment. An MRI revealed a large 6 cm×3 cm brain tumor in the leftoccipital region. The patient received resection and the pathology wasconsistent with glioblastoma. Because of her poor prognosis, the patientrefused radiation therapy or cytotoxic chemotherapy. The patient beganANP therapy in combination with a plurality of targeted agents. The ANPtherapy comprised intravenous administration of AS2-1 through asubclavian vein catheter and portable infusion pump. The dose of theAS2-1 ANP therapy was gradually increased to 40 mL (80 mg/mL) six timesdaily at 250 mL/hour (0.34 g/kg/day). Concurrently, the patient receiveddaily oral administration of dasatinib (0.7 mg/kg/day), everolimus (0.07mg/kg/day), and pazopanib (3.2 mg/kg/day). The patient also concurrentlyreceived intravenous administration of bevacizumab (10 mg/kg) every twoweeks. A baseline MRI prior to ANP therapy revealed a contrast-enhancingnodule at the margin of the postoperative cavity measuring 1.0 cm×0.6cm. A follow-up MRI after approximately 40 days of treatment indicatedthe resolution of the nodule. A subsequent MRI after approximately 75days of treatment confirmed a complete response and indicated a decreasein the postoperative cavity from 3.6 cm×3.1 cm to 2.0 cm×2.8 cm. Thepatient experienced numerous interruptions of the treatment due tosoreness in the mouth. As a result, the tumor relapsed after 103 daysand the patient died four months later.

Statements of the Disclosure Include:

Statement 1: A method for treating cancer in a patient in need thereof,the method comprising administering to a patient: (a) a plurality ofextraneous antineoplastons; and (b) a plurality of targeted agents,wherein the plurality of targeted agents comprises two or more agentsselected from the group consisting of bevacizumab, pazopanib, sorafenib,dasatinib, and everolimus.

Statement 2: A method according to Statement 1, wherein the plurality ofextraneous antineoplastons comprises phenylacetylglutaminate sodium (PG)and phenylacetylisoglutaminate sodium (iso-PG).

Statement 3: A method according to Statement 1 or Statement 2, whereinthe plurality of extraneous antineoplastons comprises about a 4:1 ratioof synthetic phenylacetylglutaminate sodium (PG) and syntheticphenylacetylisoglutaminate sodium (iso-PG).

Statement 4: A method according to Statement 2 or Statement 3,comprising intravenous administration of the plurality of extraneousantineoplastons to the patient at a dosage of from about 0.5 g/kg/day toabout 20 g/kg/day.

Statement 5: A method according to Statement 2 or Statement 3,comprising intravenous administration of the plurality of extraneousantineoplastons to the patient at a dosage of from about 2 g/kg/day toabout 8 g/kg/day.

Statement 6: A method according to any one of the preceding Statements1-3, comprising intravenous administration of the plurality ofextraneous antineoplastons in the following amounts: from about 0.4g/kg/day to about 16 g/kg/day phenylacetylglutaminate sodium (PG); andfrom about 0.1 g/kg/day to about 4 g/kg/day phenylacetylisoglutaminatesodium (iso-PG).

Statement 7: A method according to any one of the preceding Statements1-3, comprising intravenous administration of the plurality ofextraneous antineoplastons in the following amounts: from about 1.6g/kg/day to about 6.4 g/kg/day phenylacetylglutaminate sodium (PG); andfrom about 0.4 g/kg/day to about 1.6 g/kg/day phenylacetylisoglutaminatesodium (iso-PG).

Statement 8: A method according to Statement 1, wherein the plurality ofextraneous antineoplastons comprises phenylacetate (PN) andphenylacetylglutaminate (PG).

Statement 9: A method according to Statement 1 or Statement 8, whereinplurality of extraneous antineoplastons comprises about a 4:1 ratio ofphenylacetate (PN) and phenylacetylglutaminate (PG).

Statement 10: A method according to Statement 8 or Statement 9,comprising intravenous administration of the plurality of extraneousantineoplastons to the patient at a dosage of from about 0.08 g/kg/dayto about 0.6 g/kg/day.

Statement 11: A method according to Statement 8 or Statement 9,comprising intravenous administration of the plurality of extraneousantineoplastons to the patient at a dosage of from about 0.2 g/kg/day toabout 0.6 g/kg/day.

Statement 12: A method according to Statement 8 or Statement 9,comprising intravenous administration of the plurality of extraneousantineoplastons in the following amounts: from about 0.064 g/kg/day toabout 0.48 g/kg/day phenylacetate (PN); and from about 0.016 g/kg/day toabout 0.12 g/kg/day phenylacetylglutaminate sodium (PG).

Statement 13: A method according to Statement 8 or Statement 9,comprising intravenous administration of the plurality of extraneousantineoplastons in the following amounts: from about 0.16 g/kg/day toabout 0.48 g/kg/day phenylacetate (PN); and from about 0.04 g/kg/day toabout 0.12 g/kg/day phenylacetylglutaminate sodium (PG).

Statement 14: A method according to any one of the preceding Statements1-13, wherein the plurality of extraneous antineoplastons comprisesphenylacetylglutaminate sodium (PG), phenylacetylisoglutaminate sodium(iso-PG), and phenylacetate (PN).

Statement 15: A method according to any one of the preceding Statements1-14, wherein the plurality of extraneous antineoplastons isadministered intravenously.

Statement 16: A method according to any one of the preceding Statements1-15, wherein administration of the plurality of targeted agentscomprises oral administration of two or more of pazopanib, sorafenib,dasatinib, and everolimus and intravenous administration of bevacizumab.

Statement 17: A method according to any one of the preceding Statements1-16, wherein the plurality of targeted agents are administeredaccording to the following dosages: pazopanib and sorafenib isadministered orally at a dose of from about 1 mg/kg/day to about 12mg/kg/day; dasatinib is administered orally at a dose of from about 0.3mg/kg/day to about 2.0 mg/kg/day; everolimus is administered orally at adose of from about 0.03 mg/kg/day to about 0.15 mg/kg/day; andbevacizumab is administered intravenously every 1 to 3 weeks at a doseof from about 2 mg/kg/day to about 15 mg/kg/day.

Statement 18: A method according to any one of the preceding Statements1-16, wherein the plurality of targeted agents are administeredaccording to the following dosages: pazopanib and sorafenib isadministered orally at a dose of from about 2 mg/kg/day to about 6mg/kg/day; dasatinib is administered orally at a dose of from about 0.7mg/kg/day to about 1.4 mg/kg/day; everolimus is administered orally at adose of from about 0.03 mg/kg/day to about 0.10 mg/kg/day; andbevacizumab is administered intravenously every 1 to 3 weeks at a doseof from about 5 mg/kg/day to about 12 mg/kg/day.

Statement 19: A method according to any one of the preceding Statements1-18, wherein the plurality of extraneous antineoplastons and thepazopanib, sorafenib, dasatinib, and everolimus are administered dailyfor one to seven days.

Statement 20: A method according to any one of the preceding Statements1-18, wherein the plurality of extraneous antineoplastons and thepazopanib, sorafenib, dasatinib, and everolimus are administered dailyfor at least 8 weeks and the bevacizumab is administered intravenouslyevery 1 to 3 weeks for at least 8 weeks.

Statement 21: A method according to any one of the preceding Statements1-20, wherein the cancer is a glioma.

Statement 22: A method according to any one of the preceding Statements1-21, wherein the cancer is glioblastoma multiforme (GBM) or recurrentglioblastoma multiforme (RGBM).

Statement 23: A method according to any one of the preceding Statements1-22, further comprising genomic profiling of a tumor in the patient todetermine the genetic subtype.

Statement 24: A method according to any one of the preceding Statements1-23, wherein the GBM or RGBM is of the Classical subtype or theProneural subtype.

Statement 25: A method according to any one of the preceding Statements1-24, wherein the patient has failed to achieve an objective response asa result of radiation treatment and/or chemotherapy treatment, prior tothe administration of the plurality of extraneous antineoplastons andthe plurality of targeted agents.

Statement 26: A method according to any one of the preceding Statements1-25, wherein the patient has failed to achieve an objective response asa result of chemotherapy treatments comprising temozolomide (TMZ) and/orbevacizumab (BVZ), prior to the administration of the plurality ofextraneous antineoplastons and the plurality of targeted agents.

Statement 27: A method according to any one of the preceding Statements1-26, wherein the patient has undergone a resection of a glioma prior tothe administration of the plurality of extraneous antineoplastons andthe plurality of targeted agents.

Statement 28: A method for treating cancer in a patient in need thereof,the method comprising concurrently administering to a patient: (a) aplurality of extraneous antineoplastons; (b) a plurality of targetedagents, the plurality of targeted agents comprising two or more agentsselected from the group consisting of pazopanib, sorafenib, dasatinib,and everolimus; and (c) bevacizumab.

Statement 29: A method according to Statement 28, wherein the pluralityof targeted agents are administered according to the following dosages:pazopanib and sorafenib is administered orally at a dose of from about 1mg/kg/day to about 12 mg/kg/day; dasatinib is administered orally at adose of from about 0.3 mg/kg/day to about 2.0 mg/kg/day; everolimus isadministered orally at a dose of from about 0.03 mg/kg/day to about 0.15mg/kg/day; and bevacizumab is administered intravenously every 1 to 3weeks at a dose of from about 2 mg/kg/day to about 15 mg/kg/day.

Statement 30: A method according to Statement 28, wherein the pluralityof targeted agents are administered according to the following dosages:pazopanib and sorafenib is administered orally at a dose of from about 2mg/kg/day to about 6 mg/kg/day; dasatinib is administered orally at adose of from about 0.7 mg/kg/day to about 1.4 mg/kg/day; everolimus isadministered orally at a dose of from about 0.03 mg/kg/day to about 0.10mg/kg/day; and bevacizumab is administered intravenously every 1 to 3weeks at a dose of from about 5 mg/kg/day to about 12 mg/kg/day.

Statement 31: A method according to any one of the preceding Statements28-30, wherein the plurality of extraneous antineoplastons and thepazopanib, sorafenib, dasatinib, and everolimus are administered dailyfor one to seven days.

Statement 32: A method according to any one of the preceding Statements28-30, wherein the plurality of extraneous antineoplastons and thepazopanib, sorafenib, dasatinib, and everolimus are administered dailyfor at least 8 weeks and the bevacizumab is administered intravenouslyevery 1 to 3 weeks for at least 8 weeks.

Statement 33: A method according to any one of the preceding Statements28-32, wherein the plurality of extraneous antineoplastons comprisesphenylacetylglutaminate sodium (PG) and phenylacetylisoglutaminatesodium (iso-PG).

Statement 34: A method according to any one of the preceding Statements28-33, wherein the plurality of extraneous antineoplastons comprisesabout a 4:1 ratio of synthetic phenylacetylglutaminate sodium (PG) andsynthetic phenylacetylisoglutaminate sodium (iso-PG).

Statement 35: A method according to Statement 33 or Statement 34,comprising intravenous administration of the plurality of extraneousantineoplastons to the patient at a dosage of from about 0.5 g/kg/day toabout 20 g/kg/day.

Statement 36: A method according to Statement 33 or Statement 34,comprising intravenous administration of the plurality of extraneousantineoplastons to the patient at a dosage of from about 2 g/kg/day toabout 8 g/kg/day.

Statement 37: A method according to Statement 33 or Statement 34,comprising intravenous administration of the plurality of extraneousantineoplastons in the following amounts: from about 0.4 g/kg/day toabout 16 g/kg/day phenylacetylglutaminate sodium (PG); and from about0.1 g/kg/day to about 4 g/kg/day phenylacetylisoglutaminate sodium(iso-PG).

Statement 38: A method according to Statement 33 or Statement 34,comprising intravenous administration of the plurality of extraneousantineoplastons in the following amounts: from about 1.6 g/kg/day toabout 6.4 g/kg/day phenylacetylglutaminate sodium (PG); and from about0.4 g/kg/day to about 1.6 g/kg/day phenylacetylisoglutaminate sodium(iso-PG).

Statement 39: A method according to any one of the preceding Statements28-32, wherein the plurality of extraneous antineoplastons comprisesphenylacetate (PN) and phenylacetylglutaminate (PG).

Statement 40: A method according to Statement 39, wherein plurality ofextraneous antineoplastons comprises about a 4:1 ratio of phenylacetate(PN) and phenylacetylglutaminate (PG).

Statement 41: A method according to Statement 39 or Statement 40,comprising intravenous administration of the plurality of extraneousantineoplastons to the patient at a dosage of from about 0.08 g/kg/dayto about 0.6 g/kg/day.

Statement 42: A method according to Statement 39 or Statement 40,comprising intravenous administration of the plurality of extraneousantineoplastons to the patient at a dosage of from about 0.2 g/kg/day toabout 0.6 g/kg/day.

Statement 43: A method according to Statement 39 or Statement 40,comprising intravenous administration of the plurality of extraneousantineoplastons in the following amounts: from about 0.064 g/kg/day toabout 0.48 g/kg/day phenylacetate (PN); and from about 0.016 g/kg/day toabout 0.12 g/kg/day phenylacetylglutaminate sodium (PG).

Statement 44: A method according to Statement 39 or Statement 40,comprising intravenous administration of the plurality of extraneousantineoplastons in the following amounts: from about 0.16 g/kg/day toabout 0.48 g/kg/day phenylacetate (PN); and from about 0.04 g/kg/day toabout 0.12 g/kg/day phenylacetylglutaminate sodium (PG).

Statement 45: A method according to any one of the preceding Statements28-32, wherein the plurality of extraneous antineoplastons comprisesphenylacetylglutaminate sodium (PG), phenylacetylisoglutaminate sodium(iso-PG), and phenylacetate (PN).

What is claimed is:
 1. A method for treating glioblastoma multiforme ina patient in need thereof, the method comprising administering to thepatient: (a) a plurality of extraneous antineoplastons consisting of a4:1 ratio of phenylacetate (PN) and phenylacetylglutaminate sodium (PG),wherein the 4:1 ratio of PN and PG is intravenously administered to thepatient at a dosage of from about 0.08 g/kg/day to about 0.6 g/kg/day;and (b) a plurality of targeted agents consisting of bevacizumab,pazopanib, dasatinib, and everolimus wherein bevacizumab is administeredintravenously every 1 to 3 weeks at a dose of from about 2 mg/kg/day toabout 15 mg/kg/day; pazopanib is administered orally at a dose of fromabout 1 mg/kg/day to about 12 mg/kg/day; dasatinib is administeredorally at a dose of from about 0.3 mg/kg/day to about 2.0 mg/kg/day; andeverolimus is administered orally at a dose of from about 0.03 mg/kg/dayto about 0.15 mg/kg/day, wherein the administration of the plurality ofextraneous antineoplastons of (a) and the plurality of targeted agentsof (b) produces an objective response in the patient.
 2. The method ofclaim 1, wherein the method further comprises administering to thepatient an additional plurality of extraneous antineoplastons consistingof about a 4:1 ratio of synthetic phenylacetylglutaminate sodium (PG)and synthetic phenylacetylisoglutaminate sodium (iso-PG).
 3. The methodof claim 2, wherein from about 0.4 g/kg/day to about 16 g/kg/dayphenylacetylglutaminate sodium (PG); and from about 0.1 g/kg/day toabout 4 g/kg/day phenylacetylisoglutaminate sodium (iso-PG) isintravenously administered.
 4. The method of claim 1, wherein the 4:1ratio of PN and PG is intravenously administered in the followingamounts: from about 0.064 g/kg/day to about 0.48 g/kg/day phenylacetate(PN); and from about 0.016 g/kg/day to about 0.12 g/kg/dayphenylacetylglutaminate sodium (PG).
 5. The method of claim 1, whereinthe plurality of extraneous antineoplastons and the pazopanib,dasatinib, and everolimus are administered daily for at least 8 weeksand the bevacizumab is administered intravenously every 1 to 3 weeks forat least 8 weeks.
 6. The method of claim 1, further comprising genomicprofiling of a tumor in the patient to determine the genetic subtype. 7.The method of claim 6, wherein the glioblastoma multiforme is of theClassical subtype or the Proneural subtype.
 8. The method of claim 1,wherein the patient failed to achieve an objective response as a resultof at least one radiation treatment and/or at least chemotherapytreatment comprising temozolomide (TMZ) and/or bevacizumab (BVZ), priorto the administration of the plurality of extraneous antineoplastons andthe plurality of targeted agents.
 9. The method of claim 1, wherein thepatient has undergone a resection of a glioma prior to theadministration of the plurality of extraneous antineoplastons and theplurality of targeted agents.